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转人溶菌酶基因山羊奶用于胃肠道病原体的回收和治疗。

Milk from transgenic goat expressing human lysozyme for recovery and treatment of gastrointestinal pathogens.

机构信息

Laboratory of Molecular Biology and Development, University of Fortaleza, Avenida Washington Soares, 1321, Edson Queiroz, 60, 811-905 Fortaleza, Ceará, Brazil.

Laboratory of Molecular Biology and Development, University of Fortaleza, Avenida Washington Soares, 1321, Edson Queiroz, 60, 811-905 Fortaleza, Ceará, Brazil.

出版信息

Eur J Pharm Sci. 2018 Jan 15;112:79-86. doi: 10.1016/j.ejps.2017.11.005. Epub 2017 Nov 8.

Abstract

Lysozyme is an important non-specific immune protein in human milk, modulating the immune response against bacterial infections. The aim of this study was to characterize the milk of a transgenic goat expressing a recombinant human lysozyme (rhLZ) in the milk, also testing the in vitro antibacterial activity of the rhLZ milk against pathogens of the gastrointestinal tract. Milk samples collected from Tg and non-transgenic goats (nTg) from the 3rd to the 11th week of lactation were submitted to physicochemical analyses, rhLZ semi-quantification, and to rhLZ antimicrobial activity against Micrococcus luteus, Shiguella sonnei and Enterococcus faecalis. Viability and cell migration were studied in ileum epithelial cells (IEC-18) in absence or presence of E. faecalis, Staphylococcus aureus, Escherichia coli (EPEC) and S. sonnei. The expression of ZO-1 and IL-6 genes was evaluated in IEC-18 to evaluate the effect of rhLZ milk on intestinal barrier function and intestinal inflammation. Physicochemical parameters between goat Tg and nTg milk were similar and within normal values for human consumption, with hLZ concentrations being similar between Tg (224μg/mL) and human (226μg/mL) milk. The Tg milk had bactericidal activity against M. luteus, no bactericidal effect on S. sonnei, and relative to discrete sensitivity against E. feacalis than controls. Better migrating parameters were observed in cells in culture with nTg and Tg than controls. In the presence of pathogens, the Tg milk promoted improved migrating parameters than controls, except for S. sonnei, with lower cell numbers in the presence of nTg samples and E. faecalis and S. sonnei. No differences in ZO-1 relative expression patterns were observed in cultured cells, with increased expression in IL-6 in cells exposed to nTg milk than controls, with the Tg group being similar to all groups. In conclusion, goat milk containing rhLZ demonstrated valid evidence for its potential use as a nutraceutical for improvement of health and nutrition quality in humans.

摘要

溶菌酶是母乳中一种重要的非特异性免疫蛋白,可调节针对细菌感染的免疫反应。本研究的目的是表征表达重组人溶菌酶(rhLZ)的转基因山羊的牛奶,同时测试 rhLZ 牛奶对胃肠道病原体的体外抗菌活性。从泌乳第 3 周到第 11 周收集 Tg 和非转基因山羊(nTg)的牛奶样本,进行物理化学分析、rhLZ 半定量分析以及 rhLZ 对微球菌、痢疾志贺菌和粪肠球菌的抗菌活性。在不存在或存在粪肠球菌、金黄色葡萄球菌、大肠杆菌(EPEC)和痢疾志贺菌的情况下,研究了回肠上皮细胞(IEC-18)的活力和细胞迁移。评价了 rhLZ 牛奶对肠道屏障功能和肠道炎症的影响,以评估其对 IEC-18 中 ZO-1 和 IL-6 基因表达的影响。Tg 和 nTg 羊奶的物理化学参数相似,且均在人类消费的正常值范围内,hLZ 浓度在 Tg(224μg/mL)和人类(226μg/mL)奶中相似。Tg 奶对微球菌有杀菌作用,对痢疾志贺菌无杀菌作用,相对比对照物对粪肠球菌的敏感性离散。在培养物中观察到 nTg 和 Tg 细胞的迁移参数更好。在存在病原体的情况下,与对照物相比,Tg 奶促进了迁移参数的改善,除了痢疾志贺菌外,nTg 样本和粪肠球菌、痢疾志贺菌存在时细胞数量减少。在培养细胞中,ZO-1 的相对表达模式没有差异,暴露于 nTg 奶的细胞中 IL-6 的表达增加,而 Tg 组与所有组相似。总之,含有 rhLZ 的羊奶为其作为改善人类健康和营养质量的营养保健品的潜在用途提供了有效证据。

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